Process for the preparation of malted cereals

ABSTRACT

Proces for the preparation of malted cereals, wherein the steeping step includes one or more wetting stages at a temperature and 30° C. preferably between 10 and 20° C., until the material has a moisture content between 20 and 60% by weight, preferably between 38 and 47%, wherein aftr a germination period between 2 and 7 days, preferably between 3 to 6 days at a temperature between 10 and 30° C., preferably between 14 and 18° C., the steeped and germinated cereals are preferably kilned by increasing the temperature to values between 40 and 150° C. until the material has a moisture content between 2 and 15% by weight, and wherein one or more microbial cultures selected from the group consisting of one or more bacteria and/or one of more activated spores are added in one or more times during the process.

FIELD OF THE INVENTION

[0001] The present invention is related to an improved pxoess for tepreparation of malted corealws, the improved malted cereals obtained andtheir use, especially In biotechnological processos for the preparatonof beverages.

TECHNOLOGICAL BACKGROUND OF THE INVENTION

[0002] Cereals, such as barley, wheat, rye, corn, oats, rice, millettriticale, and sorghum, are used for the production of beverages. Inmost cases, they have been subjected to a malting process to takeadvantage of their increased enzymatic potential.

[0003] In traditional malting processes, the moisture content of cerealsis raised either by immersion(s) andfor sproying(s), and the resultinghigh-moisture content cereal is allowed to germinate. After reaching theproper physological condition, it is preferably subrnitted to (a) dryingstep(s). In what follows, the term steeping refers to the increase inmoisture level, while the term germination is used in the way it is inplant physiology. The drying operations are referred to as kilning andthe term malting involves all operations needed to convert barley (orother cereals) to barley malts (or other cereal malts).

[0004] The quality of the malt obtained is, to a large extent,determined by the presence of plant endogenous enzymes genereted duringthe melting process. For instance, with cereals like barley used as araw materal for the malt production. the variety, the oomposwon of themicrobal flora and the environmental factors, such as agriculturalpractice, Influence the quality of the malt. During cultivation andstorage, cereals are contaminated with bacteria and fungi. In themelting plant, neither the air, the water, nor the equipment aresterile, and the conditions of humidity, pH and temperature favor thegrowth of the microbial populations. To improve the quality of maltedcereals, such as barley, enzymes have been added to the malted cereal.

[0005] The variable cereal quality and the lack of means to make up fordeficiencies during the malting process result in variability in maltquality and enzymatic activity. In many instances, this has to do withan imbalanced of specific enzymatic potential and insufficient cell walldegradation. Apart from this, problems with microbial safety can oocur.As a consequence of the defects In malt, quality problems occur in theproduction of beer, such as a poor filtration of the wort.

STATE OF THE ART

[0006] During the malting of cereals, the micrflora present on thecereals develops and the quality of the matt and beverages is influencedby the activity of the ister microorganisms.

[0007] In analogy with other blotechnological processes, there have beenattempts to optimize malt quality aspects by the addition of startercultures during the malting process (Boivin, P. & Malanda, M., Influenceof Starter Cultures in Malting on the Microflora Development and MattQuality, EBC, Proceedings of the 24th Congress, pp. 95-102 (1993);Haikara, A. et al., Lactic Starter Cultures in Malting - A NovelSolution to Gushing Problems, European Brewery Convention, Proceedingsof the 24th Congress, pp. 163-172 (1993)).

[0008] Addition of spores of Geotrichum candidum to the steeping waterresults in the inhibition of the development of undesirablemicroorganisms and in a decrease of the filtration time of wort made ofthe obtained malt. Treatment with Geotrichum candidum also inhibits theformation of mycotoxins by Fusarium app.

[0009] The influence of Lactobacillus plantarum and Pediocoscuspentosaceus has been tested on the microflora during malting, and It hasbeen found that these cultures act as natural preservations as theyrestrict the growth of Fusarium and prevent gushing.

[0010] The Internatonal patent application WO 94/29430 describes aprocess for improving the properties of malted cereals wherein startercultures which comprise moulds, yeasts or bacteria are added priorand/or during malting of said cereals.

[0011] The preferred bacteria used are lactic acid producing bacteria,such as various Lactobacilli, e.g., Lactobacillus casel, Lactobacilluscasel var. rarmnosus, Lactobacillus fenmentum, Laciobacillus plantarumand Lactobacllus brevis, and bacteria of the genus Pediococcus, e.g.,Pediococcus acidilactici.

[0012] Preferred moulds are moulds of the genus Aspergillus andGeotrichum, like Geotrichum candidum.

[0013] The international patent application WO94/16053 describes aprocess for treating cereals for inhibiting growth of unwanted microbialspecies by inoculating the cereals during the germination process with alactic acid bacteria preparation or a preparation produced by lacticacid bateria. The preferred bacteria are lactic acid bacteria belongingto genus Lactococcus, Leuconostoc, Pedlococcus or Lactobacillus.

[0014] The British patent application GB-1211779 provides a method forthe automatic control and regulation of a malting process. It enablesone to determine the parameters necessary for a successful automaticallycontrolled and regulated malting process.

[0015] In the proceedings of the European Brewery Convention, volume 16,1977, pages 245 to 254, the influence of some fungi on malt quality isdescribed, more speciffcally, contamination of barley malt wfth fungiwhl has led to gushing and other qualtative changes In the beer.

[0016] The German patent application DE-3028360 disclosm a rnetod tomake malt out of corn.

[0017] However, malt prepared according to the present invention is ofbetter quality than that prepared according to WO 94/29430. This isexemplified by higher β-glucanase and xylanase activities, lower -glucancontents in malt and wort and improved European Brewery Conventionanalytical data.

AIMS OF THE INVENTION

[0018] The present invention aims to provide an improved preparedprocess for malted cereals and improved malted cereals.

[0019] A main aim of the invention is to provide an improved preparationprocess for matted cereals and improved malted cereals in terms ofbrewing performances, especially malted cereals having an improvedquality in terms of enzymatic potential and microbial safety.

[0020] Another aim is to provide a process and improved malted cerealswhich vary less in quality with the raw material used.

[0021] A further aim of the invention is to obtain malted cereals whichimprove the biotechnological production process of beverges and mayimprove the properties of the said obtained beverages.

SUMMARY OF THE INVENTION

[0022] The present invention is directed to a process for thepreparation of a malted cereal, the malted cereal which is the productof the proess of the invention, and a combination of wetted or moistenedcereal and activated spores which, when held for a sufficient time andtemperature, provide a malted cereal product of enhanced enzymaticactivity. The product of the invention has enhanced enzymatic activityof at least one enzyme, such as β-glucanase, xylanase, amylase,naturally occurring enymes, and/or protease activity, over malted cerealproducts which are similarly prepared with wetted cereal products withor without microorganisms.

[0023] The prss of the invention utilizes activated spores frommicroorganisms such as bacteria or moulds. The process generallycomprises combining water, the cereal and activated spores and holdingthe combination until a malted cereal of enhanced enzymatic activity isformed. Generally the combination is made by inoculating the moistenedcereal with the activated spores, but the activated spores and cerealmay be combined before or after the moistening of the cereal. In theprocess of the invention, the combination of weted cereal and activatedspores has a concentration of activated spores, holding time and holdingtemperature which are effective for providing the malted cereal with anincrease In enzymatic acity of at least one enzyme, such as tlucanase,xyanase, amylase, naturally occurring enzymes, and/or protease activity,whith is greater than the enzymatic activity which is obtained byholding the wetted oreal without activated spores, or even with thebacteria or moulds from which the spores come.

[0024] In an important aspect, the cereal, activated spores are combinedbefore or after the time of wetting the cereal and the combination isheld at a temperature of at least about 5° C. and not more than about30° C., preferaby between about 10° to about 20° C. and the activatedspores are at a concentration In the combination to obtain an increasein enzymatic activity of the malted cereal. In another important aspect,the wetted or moistened cereal and activated spore oombinain is held fora time and temperature until the cereal has a moisture content of atleast about 20 weight percent. In yet another aspect, after the wettedcereal has attained an increased moisture content and has started togerminate, it is dried to a moisture content of not more than about 15weight percent. In still another important aspect, the moistened cerealand activated spores are held together until the cereal has a moisturecontent of btween about 20 to about 60 weight percent, preferably fromabout 38 to about 47 weight percent, and has germinated for about 2 toabout 7 days, preferably about 3 to about 6 days, at a temperature offrom about 10° to about 30° C., preferably from about 14° to about 18°C. In important aspect, the germinated cereal is dried at a temperatureof from about 40° to about 150° C.. preferably beteen about 45° and 85°C. until the dried malted cereal has a moisture content of from about 2to about 15 weight percent moisture, preftrably from about 4 to abut 7weight percent moisture. Importantly, the process of the invention mayincrease β-glucanase activity of a malted cereal by a factor of at leastabout 4 as compared to a matted cereal prepared without activated sporesaccording to the invention.

[0025] In another important aspect, the present invention provdesd amalted cereal having a higher quantity of acrospire lengths that weresignificantly longer in comparison to acrospire lengths when traditionalmalting methods were used.

[0026] In an important aspect, the cereals which may be used in theinvention include barley, wheat and sorghum. In one aspect of theinvention the spores of moulds are used. In another aspect, the sporesof bacteria are used.

[0027] In another aspect, the oprals may be disinfected or may not bedisinfected.

[0028] Preferably, for the preparation of matted barley, the spores frombacteria are from gram positive bacteria or gram negative bacteria,selected from the group of Micrococcus spp., Streptococcus spp.,Leuconostoc spp., Pediococcus spp. preferentially Pediococcushalophilus, Pediococcus cerevisiae, Pediococcus damnosus, Pediococccushemophilus, Pediococccus parvulus, Pediococcus soyae, Lactococcus spp.,Lactobacillus spp. preferentially Lactobacillus acidophilus,Lactobacillus amylovorus, preferentially Lactobacellus amylovorus strainATCC 33620, Lactobacillus bavaricus, Lactobacillus bifermentans,Lactobacillus brevis var llndneri, Lactobacillus casei var casel,Lactobacillus delbrueckii, Lactobacillus delbrueckii var iacts,Lactobacillus delbrueckii var bulgaricus, Lactobacillus fermenti,Lactobacillus gasserii, Lactobactilus helveticus, Lactobacillushilgardii, Lactobacillus renteril, Lactobacillus sake, Lactobacillussativorius, Lactobacillus cremoris, Lactobacillus kefir, Lactobacilluspentoceticus, Lactobacillus cellobiosus, Lactobacillus bruxellensis,Lactobacillus buchneii, Lactobacillus coryneformis, Lactobacillusconfusus, Lactobacillus floretinus, Lactobacillus viridescens,Corynebactrium spp., Propionibacterium spp., Bifidobaterium spp.,Streptomyces spp., Bacillus spp., preferentially Bacillus subtilis stainATCC 6051, preferentially Bacillus circulans, Sporolactobacillus app.,Acetobacter spp., Agrobacterlum spp., Alcaligenes spp., Peoudomonasspp., preferentially Pseudomonas amylophilia, Pseudormonas aeruginosa,Pseudomonas cocovenenans, Pseudomonas mexdcana, Pseudomonas pudomaliel,Gluconobadler spp., Enterobacter spp., Erwinia spp., Kiebsiella spp.,and Proteus spp.

[0029] Preferably, for the preparation of matted barley spores are fromfungi which are selected from the group (genera as described byAinsworth and Bisby's dictionary of the fungi, 8th edition, 1995, editedby DL Hawkworth, PM Kirk, BC Sutton, and DN Pegler (632 pp) CabIntermal) of Ascomycota preferentially Dothideales, preferentiallyMycosphaerellaceae preferentially Mycosphaerell spp., Ventunaoaepreferentially Venturia spp. Eurotleles preferentially Monascaceaepreferentially Monascus spp., Triciococcmacese preferentially Emericillaspp., Euroteum spp., Eupenicillium spp., Neosartorya spp., Talaromycesspp., Hypocreales preferentially Hypocrecase preferentially Hypocreaspp., Saccharomycetales preferentially Dipodascacae, Dipodascus spp.,Galactomyos spp., Endomycetaceae preferentially Endomyces spp.,Metschnkowiaceae preferentially Guilliermondella spp.,Saccharomycetaceae preferentially Debaryomyces spp., Dekkera spp.,Pichis spp., preferetially Pichia anomala, preferentially Pichia anomalastrain ATCC 8168. Kluyveromyces spp. Saccharomyces spp., Torulasporaspp., Zygosaccharomyces spp., Saccharomycodaceae preferentiallyHanseniaspora spp.; Schizosaccharomycetales preferentiallySchlzosaccharomycetaceae preferentially Schizosaccharomyces spp.,Sordariales preferentially Chaetomisse, Chaotomium spp., preferentiallyChaetomium vireacens strain ATCC 32319, Sordariacea preferentiallyNeurospora spp., Zygomycota preferentially Mucorales preferentiallyMucoraceae preferentially Absidia spp., Amylomyces spp., Rhizomucorspp., Actinomucor spp., Thermomucor spp., Chiamydomucor spp., Mucor spp.preferentially Mucor circinelloides, Mucor grisecyanus, Mucor hiemalis,Mucor indicus, Muoor mucedo, Mucor piriformis, Mucor plumbous, Mucorpraini, Mucor pusillus, Mucor silvaticus, Mucor javanicus, Mucorracemosus, Mucor rouxianus, Muoor rouxil, Mucor aromaticus, Mucorflevus, Mucor miehei, Rhizopus spp. preferentially Rhizopus arrhizus,Rhizopus oligosporus, Rhizopus oryzae preferentially Rhizopus oryzaestrain ATCC 4858, Rhizopus oryzae strain ATCC 9363, Rhizopus oryzaestrain NRRL 1891, Rhizopus oryze strain NRRL 1472, Rhizopus stolonifer,Rhizopus thailandensis, preferentially Rhizopus thallandensis strainATCC 20344, Rhizopus formosaensis, preferentially Rhizopus formosaensisstrain ATCC 26612, Rhizopus chinensis, Rhizopus cohnii, Rhimpusjaponicus, Rhizopus nodosus, Rhizopus delemar, Rhizopus acetorinus,Rhizopus chimydosporus, Rhizopus circinans, Rhizopus javanicus, Rhizopuspeka, Rhizopus saito, Rhizopus triticl, Rhizopus niveus, Rhizopusmicrosporus, Mitosporic fungi preferentially Aureobasidium spp.,Acremonium spp., Cercospom spp., Eplcobcum spp., Monlila spp.preferentially Monilia candida, Monilia sitophila, Mycoderma spp.,Candida spp., preferentially Candida diddenslae, Candida edax, Candideetchellsii, Candida kefir, Candida krisel, Candida lactose, Candidalambica, Candida mlinil, Candida utills, Candida milleri, Candidamycoderma, Candida parpilosis, Candida obtux, Candida troploale, Candidavalida, Candida verutllis, Candida guilliermondii, Rhodotmlea spp.,Torulopsis spp., Geotrichum spp. preferentially Geotrichum amycelium,Geotrichum armillariae, Geotrichum asteroides, Geotrichum bipunctatum,Geotrichum dulcitum, Geotrichum eriense, Geotrichum fici, Geotrichumflavo-brunneum, Geotrichum fragrans, Geotrichum gracile, Geotrichumheritum, Geotrichum kiebaknii, Geotrichum penicillatum, Geotrichumhirtum, Geotrichum pseudocandidum, Geotrichum rectanguistum, Geotrichumsuaveolens, Geotrichum vanryiae, Geotrichum loubieri, Geotrichummicrosporum, Ciadosporium spp., Trichoderma spp. preferentiallyTrichoderma hamatum, Trichoderma harzianum, Trichoderma koningii,Trichoderma pseudokoningli, Trichoderma resei, preferentiallyTrichoderma resei strain ATCC 5875, Trichoderma virgatm, Trichodermaviride, Oldium spp., Altemaria spp. preferentially Alternaria alternata,Alternaria tenuis, Helminthosporium spp. preferentially Helminthosporiumgramineum, Helminrium sativum, Helminthosporium teres, Aspergilius spp.preferentially Aspergillus ochraseus Group, Aspergillus nidulans Group,Aspergglus vemicolor Group, Aspergillus wentii Group, Aspergilluscandidus Group, Aspergillus flavus Group, Aspergillus niger Group,Aspergillus oryzae strain ATCC 14156, Penicillum spp. preferentiallyPenicillum aculeatum, Penicillum citrinum, Penlcillium claviforme,Penicillum funiculosum, Penicillum italicum, Penicillum lanoso-viride,Penicillum emersonil, Peniclilum lilacinum, Penicillum expansum, andmixtures thereof.

[0030] Preferably, for the preparation of malted cereals other thenmalted barley, especially for the preparation of malted wheat, rye,corn, oats, rice, millet, triticale, and sorghum, said bacteria are grampositive or gram negative bacteria selected from the group ofMicrococcus spp., Streptococcus spp., Leuconostoc spp., Pediococcusspp., Lactococcus spp., Lactobacillus spp., Corynebacterum spp.,Propionibacterium spp., Bifidobacterium spp., Streptomyces spp.,Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacteriumspp., Alcaligenes spp., Pseudomonas spp., Gluconobacter spp.,Enterobacter spp., Erwinia spp., Klebsiella spp., Proteus spp. or amixture thereof; and said fungi are fungi selected from the group of:Ascomycota preferentially Dothideales preferentially Mycophaerellaceaepreferentially Mycosphaerella spp., Venturiaceae preferentially Venturaspp.; Eurotiales preferentially Monascaceae preferentially Monascusspp., Trichocomaceae preferentially Emericilla spp., Euroteum spp.,Eupenicillium spp., Neosartorya spp., Talaromyces spp., Hypocrealespreferentially Hypocreaceae preferentially Hypocrea spp.,Saccharomycetales preferentially Dipodascaceae preferentially Dipodascusspp., Galactomyces spp., Endomycetaceae preferentially Endomyces spp.,Metschnikowiaceae preferentially Guilliermondella spp.,Saccharomycetaceae preferentially Debaryomyces spp., Dekkera spp.,Pichia spp., Kluyveromyces spp., Saccharomyces spp., Torulaspora spp.,Zygosaccharomyces spp., Saccaromycodaceae preferentially Hanseniasporaspp., Schizosaccheromycetales preferentially Schizosaccharomycetaceaepreferentially Schlzosaccharomyces spp.; Sordariales preferentiallyChaetomiaceae preferentially Chaetomium spp., Sordariacesepreferentially Neurospora spp., Zygomycota preferentially Mucoraiespreferentially Mucoraceae preferentially Absidia spp., Amylomyces spp.,Rhizomucor spp., Actinomucor spp., Thermomucor spp., Clamydomucor spp.,Mucor spp., Rhizopus spp.; preferentially Rhizopus oryza strain ATCC9363. Mitosporic fungi preferentially Aureobasidium spp., Acremoniumspp., Cerocospora spp., Epicoccum spp., Monilia spp., Mycoderma spp.,Candida spp., Rhodotorula spp., Torulopsis spp., Geotrichum spp.,Cladosporium spp., Trichoderma spp., Oldium spp., Alternaria spp.,Helminthosporium spp., Aspergillus spp., Penicillium spp.

DEFINITIONS

[0031] As used herein the term “spore” refers to a dormant and highlyresistant reproductive cell fomned by bacteria and fungi in response toenvironment conditions that do not favor the growth of the organism.When exposed to favorable environmental conditions, spores are capableof developing Into a viable adult organism without fusion with anothercell.

[0032] As used herein the term “activated spore” means a spore havingone of the following properties:

[0033] i) The spore is swollen such that its size increase by a factorof between about 1.2 and about 10 over its dormant size; and/or

[0034] ii) one or more germ tubes per spore is formed.

[0035] Activated spores are prepared by one or a combination of thefollowing treatments.

[0036] i) cycles of wetting and/or drying;

[0037] ii) addition of appropriate nutritional supplies (such as anitrogen source, preferably amino acids and/or a carbon source,preferably mono- or disaccharides) or spore elements;

[0038] iii) exposure to temperature changes, preferably within atemperature range of about oe to about 80° C..

[0039] iv) exposure to changes in pH, preferably within a pH range ofabout 2.0 to about 8.0, more preferably about 3.0 to about 6.0.

[0040] The term “germination” as used herein means the beginning orresumption of growth by a seed. In accordance with the process of thepresent invention, germination begins to occur during and/or after thecereal has been steeped. Germination of cereals is generally understoodto mean hydration of the seed, swelling of the cereal and inducinggrowth of the embryo. Environmental factors affecting germinationinclude moisture, temperature and oxygen level. A rapid increase incells of the root stem leads to root development, while correspondinggrowth sends forth a shoot.

[0041] As used herein, the term “steeping” refers to weing of thecereal. Wetting may include one or more stages over a time andtemperature effective for providing a moisture content of between about20% and about 60% by weight.

[0042] The term “specific ac” as used herein refers to the concentrationand activity of an enzyme in a preparation. The specific activity of apreparation is reported as units/mg protein. One unit of enzyme is thatamount that catalyzes the formation of 1 mmole of product per minuteunder defined conditions. The amount of enzyme present in a preparationis measured using standard protein assay techniques and catalyticactivity is determined by following the formation of product or removalof substrate over time.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0043] According to a prefrd embodiment, the preparatbn process ofmalted cereals according to the invention comprises the following steps:a steeping step includes one or more wetting stages or the total time ofsubmersion in water during steeping for physiological reasons does notexceed 30 hours (preferably 10 to 25 hours) or the kilning step includesmore then two temperatus steps and the microbial cultures which areadded, are preferably selected from the group consisting of Rhizopusspp., preferably Rhizopus oryzae, such as Rhizopus oryze strain ATCC9363and/or Pseudomonas spp., prefably Pseudomonas herbicola.

[0044] According to the invention, the malted cereals are selected fromthe group of barley, wheat, rye, corn, oats, rice, millet, trficoale,sorghum and the like.

[0045] In the process according to the invention, the same or deferentactivated spores are added in one or more time(s). The use of activatedspores greatly enhances their contribution to improved malt quality,most likely because of more vigorous growth. The activated spores haveone of the following properties: the treated spores are significantlymore swollen than their dormant, size, more particularly, the size ofthe spores is increased by a factor preferably between 1.2 and 10 overtheir dormant size and/or one or more germ tubes per spore are formed.The activated spores are prepared by subjecting them to environmentalchanges, preferably by one or a combination of the following treatments;

[0046] (a) cycles of wetting and/or drying;

[0047] (b) addion of appropriate nutritional supplies (such ass anitrogen source, preferably amino acids and/or a carbon source,preferably mono- or disaccharides) or spore elements;

[0048] (c) exposure to temperaure changes, prsftmbly within a tempemwurerange of 0° to 80° C.;

[0049] (d) exposure to changes in pH, preferably wfthln a pH range of2.0 to 8.0, more pmftrably between 3.0 and 6.0.

[0050] The activated spores may be introduced before or during themalting process. For example, the activated spores may be introducedduring the various malting or steeping stages before or after immersionof the cereal.

[0051] The concentration of the spores may vary depednding on theconditions of the malting process and the type of active spore beingutilized. Generally about 1×10² to about 1×10⁷, preferably about 1×10³toabout 1×10⁵ activated spores per gram air dry cereal is utilized.

[0052] The present invention also concerns the malted cereals obtainedaccording to the process of the invention, which present improvedEuropean Brewery Convention analysis results. Said improvements may haveto do with modification and/or increased hydrolytic enzyme activies. Atthe same time, a decreased level of toxins, an increased microbialsafety by e.g., outcompeting undesirable microbial flora such asFusarium and/or an increased acceptability compared to the maltedcereals according to the state of the art, may be observed.

[0053] For instance, the malted cereals acoording to the invention mayhave a lower β-glucan content or a higher enzyme activw such as, forexample, β-glucanase or xylanase actvy (as represented in the folowingexamples and figures) than the malted cereals according to the state ofthe art. This allows for a better processability of the malt in wort andbeer production as exemplified by increased rates of filtration.

[0054] The activated spores and cereal may be combined and wetted bysubmersion in water to steep the combinatlon which should not exceed 30hours. The activated spores can also be sprayed on the barley during thesteep period or during the germination process. The pH during thisperiod should be from about 1.5 to about 14, preferably about 4 to about6. β-glucanase activity of malted barley made according to the inventionis higher than 700 units/kg and xylanase activity is higher than 250units/kg.

[0055] An object of the present invention concerns the use of the maltedcereals according to the invention for the preparation of beverages.

[0056] The invention is also related to these improved beverages.

[0057] The improved malted cereals according to the inventon could alsobe used in other biotechnological processes well known by the PersonSkilled in the Art, in which in most cases advantage is taken of theirimproved quality.

[0058] The present invention will be further described in variousexaerles in view of the following drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

[0059]FIG. 1 represents the β-glucanase activity of malted barleyobtained according to the preparation process of example 1. (legend: seeexample 1).

[0060]FIG. 2 represents the xylanese activity of malted barley obtainedaccording to the preparation process of example 1. (legend: see example1).

[0061]FIG. 3 represents the β-glucanase activity of malted barleyobtained according to the preparation process of example 3. (legend: seeexample 3).

[0062]FIG. 4 represents the xylanase activity of malted barley obtainedaccording to the preparation prooess of example 3. (legend: see example3).

[0063]FIG. 5 represents the relative increase factory (R.I.F.) forbacterial populations (see text, malt evaluation, example 2) (legend:see example 2).

EXAMPLE 1

[0064] 1. Preparation of Microbial Cultures

[0065] Strain

[0066] S46: Rhizopus oryzae ATCC 9383.

[0067] Preparation of the Spore Suspension

[0068] the strain was grown on PDA (Potato Dextrose Agar, Oxoid) forapproximately 10 days at 28° C.;

[0069] the spores were harvested by flooding the cultures with sterilephysiological saline (0.9% NaCl) and by rubbing the sporulated myceliumgently with a sterile spatula;

[0070] the spore suspension was washed twioe with sterile physiologicalsaline (0.9% NaCl) by centrifugation (5500 rpm, Sorvall type SS-34®, for15 min.) and resuspended in sterile physiological saline (0.9% NaCl);

[0071] the spore density was determined micrampically using a Thomacounting chamber.

[0072] Activation of the Spore Suspension

[0073] 10⁷ spores were transferred into 20 ml of sterile, acidifed TSB(Tryptic Soy Broth, Oxoid), pH=4.0 and incubated in a shaking water bathduring 5 to 6 hours at ±42° C.;

[0074] The activated spores were harvested by centrifugation (3500 rpm,Sorvall type SS-34®, for 15 min.), washed once with sterilephysiological saline (0.9% NaCl) by centeugallon (3500 rpm, Soivall typeSS-34®, for 15 min.) and resuspended in sterile physiological saline(0.9% NaCl).

[0075] 2. Barlay

[0076] Plaisant - 1994 French harvest.

[0077] 3. Process.

[0078] Setup

[0079] Malts were made by four different malting processes:

[0080] A1. traditional malting

[0081]  (without inoculation of any spore suspension)

[0082] B1. malting process using non-activated spores

[0083]  (inoculation of the stewp barley with a suspension ofnon-activated spores of Rhizopus oryzae ATCC 9363)

[0084] C1. malting process according to the invention

[0085]  (inoculation of fte seped barley with a suspension of activatedspores of Rhizopus oryzae ATCC 9363)

[0086] D1. malting process according to the inverion

[0087]  (inoculation of the steeped barley during the first wet stagewith a suspension of activated spores of Rhizopus oryzme ATCC 9363)

[0088] Steeping

[0089] the steeping was carried out on a 2 kg base with a total water(tap water) to air dry barley ratio of 1.5:1;

[0090] use was made of 2 fermentors (Biofio III, New BrunswickScientific), in which perforated plates were placed;

[0091] temperature was only controlled during the wet stages, during theair rest stages, the system was allowed to reach room temperature (±20°C.);

[0092] during the whole steeping period, the barley was serated (4 litersterile air per minute);

[0093] steeping was carried out by immersion using the following scheme;Temperature (° C.) Duration (h) First wet stage 13 6:00 First air reststage 20 17:00  Second wet stage 14 5:00 Second air rest stage 20 15:30 Third wet stage 16 2:30

[0094] Addition of the Microbial Cultures

[0095] ±480 g of steeped barley was immersned in 0.5 liter of tap Waterwhich contained no spores (A1), non-activated spores of Rhizopus oryzaeATCC 9363 (B1) or activated spores of Rhizopus oryzae ATCC 9363 (C1,according to the invention); for B1 and C1, the steeped barley wasinoculated with 10⁴ spores per gram of air dry barley;

[0096] during the steeping, 10⁴ activated spores per gram air dry barleywere inoculated to the water of the first wet stage (D1);

[0097] the fluid was removed by draining.

[0098] Germination

[0099] germination was carried out in a cylindrical container withperformated lids at a temperature of 16°-18° C. during 4 days;

[0100] air was supplied by natural diffusion;

[0101] the containers were slowly rotated on an electrically controlledroller system (Cellroll®, Tecnorama); i.e., every two hours thecontainers were rolled for 15 min. at 1 rpm.

[0102] Kilning

[0103] the kilning was carried out in a Joe White malting unit(Australia), Air flow Recirc. Temp. Durat. (%) Air (%) (° C.) (h) Firstkilning stage 25 0 62 3:00 Second kilning stage 25 0 65 2:00 Thirdkilning stage 25 0 68 2.00 Fourth kilning stage 25 25 73 2:00 Fifthkilning stage 25 50 78 1:00 Sixth kilning stage 25 75 80 2:00 Seventhkilning stage 25 100 83 6:00 Shut down air off Time out

[0104] 4. Methods of Analysis and Results

[0105] Methods for determination and units of moisture, extract, extractdifference, color, total protein content, soluble protein content,Kobach index, pH, diasttic power, according to Analytica-EuropeanBrewery Convention (Fourth Edition, 1987, Brauerei undGetränke-Rundschau).

[0106] Metods for determination and units of turbidity, frability,homogeneity, whole grains, b-glucan content, according toAnalytica-European Brewery Convention (Fourth Edition, 1987, Brauereiund Getränke-Rundschau, supplement published in 1989).

[0107] Postcoloration of the wort is determined after boiling thecongress wort under reflux at 108° C. during 2 hours.

[0108] The viscosity of the congress wort is determined with theDelta-viscosimater.

[0109] For the determination of the filtration volume, the congress wortis filtered over a Schleicher and Schuell 597½folded filter. The volume(in mil) that is obtained after 1 hour of filtration is the filtrationvolume of the wort.

[0110] Modification is determined with the Calcofluor apparatus(Haffmans) according to the Carisberg method (Analytica-Europeon BreweryConvention (Fourth Edition, 1987, Brauerei und Getränke-Rundschau).

[0111] The β-glucanase and xylanase activities are determined with theβ-glucazym method (Megazme (Austr.) Pty Ltd. (April, 1993) and thexylazym method ((Megazyme (Austr.) Pty Ltd. (September 1995)),respectively. Malting Malting Malting Process Process ProcessTraditional using non- according according Malting activated to the tothe Process spores Invention Invention (A1) (B1) (C1) (D1) Moisture 3.94.1 3.8 4.3 Extract 80.3 80.4 80.3 79.8 Extract 0.8 0.8 0.4 1.1Difference Color 3.3 3.3 4.1 4.1 Wort Turbidity 1.3 1.2 0.7 0.8Postcoloratlon 6 6 7.3 7.5 Total Protein 10.1 10.3 10 10.1 ContentSoluble Protein 4.1 4.4 4.8 5.2 Content Kolbach Index 40.6 42.7 48 51.0Viscosity 1.57 1.52 1.52 1.54 pH 6.05 6.3 5.87 5.79 Diastatic Power 345349 352 419 Whole Grains 0.3 0.3 0.1 ND Friability 83 82 83.9 NDHomogeneity 98.5 97.9 98.6 ND β-glucan content 122 108 46 <40 FiltrationVolume 210 265 290 275 Modification 88.2 90.5 93.4 ND β-glucanase 214371 683 3656 Activity Xylanase Activity 28 34 56 984

[0112]FIGS. 1 and 2 represent the β-glucanase and xcylanase activity,respectively of the obtained malted barley (A1, B1, C1, D1). Thesemalted barleys are obtained according to a traditional malting process(A1) or using non-activated spores during the malting process (B1) oraccording to the above-described malting process of the invention (C1,D1). The β-glucanase activity was determined with the β-glucazym method(Megazyme (Austr.) Pty Ltd. (April, 1993)). Therefore, malt β-glucanaseactivity (U/kg) was calculated at 380×E (590 nm)+20. The xylanaseactivity was determined with the endo 1-4-xylazym method (Megazyme(Austr.) Pty Ltd. (September 1995)). Therefore, malt xylanese actvity(U/kg) was calculated as (46.8×E (590 nm)+0.9)×5).

EXAMPLE 2

[0113] 1. Preparation of Microbial Cultures

[0114] Strain

[0115] S46: Rhizopus oryzae ATCC 9363.

[0116] as described in Example 1.

[0117] Preparation of the Spore Suspension

[0118] as described in Example 1.

[0119] Activation of the Spore Suspension

[0120] as described in Example 1.

[0121] 2. Barley

[0122] Stander - 1995 North American harvest.

[0123] 3. Process

[0124] Setup

[0125] Malts were made by six different malting processes:

[0126] A2, tradnonal malting process

[0127]  (without inoculation of any spore suspension)

[0128] B2, malting process using non-activated spores

[0129]  (inoculation of the steeped barley with a suspension ofnon-activated spores of Rhizopus oryzae ATCC 9363)

[0130] C2, malting process according to the invention

[0131]  (inoculation of the steeped barley during the first wet stagewith a suspension of activated spores of Rhizopus oryzae ATCC 9383)

[0132] D2, malting process according to the invention

[0133]  (inoculation of the steeped barley during the second wet stagewith a suspension of activated spores of Rhizopus oryzae ATCC 9363)

[0134] E2, malting process according to the invention

[0135]  (inoculation of the steeped barley during the third wet stagewith a suspension of activated spores of Rhizopus oryzae ATCC 9363)

[0136] F2. malting process according to the invention

[0137]  (inoculation of the steeped barley with a suspension ofactivated spores of Rhizopus oryzae ATCC 9363)

[0138] Steeping and Addition of the Microbial Cultures

[0139] the steeping was carried out on a 300 g base with a total water(tap water) to air dry barley ratio of 5:3;

[0140] use was made of 2000 ml flasks;

[0141] a tempertre of 18° C. was maintained during the wet stages andduring the air rest stages;

[0142] during the whole steeping period, the barley was aerated by meansof compressed air;

[0143] steeping was carried out by immersion using the followingschedule; Duration (h) First wet stage 6:00 First air rest stage 18:00 Second wet stage 5:00 Second air rest stage 19:00  Third wet stage 2:00

[0144] during the steeping, 10⁴ activated spores per gram of air drybarley were inoculated to the water of the first wet stage (C2), of thesecond wet stage (D2) or of the third wet stage (E2) before immersion ofthe barley;

[0145] the steeped barley was immersed in 0.5 liters of tap water whichcontained no spores (A2), non-actvated (B2) or activated (C2, D2, E2,F2) spores;

[0146] for B2, and F2, the steeped barley was inoculated with 10⁴ sporesper gram of air dry barley,

[0147] the fluid was removed by draining.

[0148] Germinaton

[0149] as described in Example 1.

[0150] Kilning

[0151] as described in Example 1.

[0152] Malt evaluation

[0153] Determination of the Increase of the Bacterial Population

[0154] To judge the evolution of the bacterial population during themalting process, a relative increase factor (R.I.F.) was determined bydividing the total bacterial count occurring on the green malt by thetotal bacterial count occurring on the barley. The total bacterial countwas determined after plating appropriate dilutions of an extract of thekernels on Tryptic Soy Agar (Oxoid) supplemented with 100 ppmpirnaricine and after incubation at 28° C. for 3 days.

[0155]FIG. 5 shows the increase of the bacterial population during themalting according to the preparation process of Example 2.

EXAMPLE 3

[0156] 1. Preparation of Microbial Cultures

[0157] Strain

[0158] S 46: Rhizopus oryzae ATCC 9363

[0159] Preparation of the Spore Suspension

[0160] as described in Example 1

[0161] Activation of the Spore Suspension

[0162] as described in Example 1

[0163] 2. Barley

[0164] Plaisant - 1994 French harvest;

[0165] 3. Process

[0166] Setup

[0167] Malts were made by three different malting processs:

[0168] A3. traditional malting

[0169]  (without inoculavon of any spore suspension)

[0170] B3. malting process using non-activited spores

[0171]  (inoculation of the steeped barley with a suspension ofnon-activated spores of Rhizopus oryzae ATCC 9363)

[0172] C3. malting process according to the invention

[0173]  (inoculation of the steeped barley with a suspension of activtedspores of Rhizopus oryzsa ATCC 9363)

[0174] Steeping

[0175] the steeping was carried out on a 2 kg base air dry barley with atotal water (tap water) to air dry barley ratio of 1.5:1;

[0176] the pH of the steeping water was controlled at pH=5.5 by additionof lactic acid and NaOH;

[0177] a fermentor (Bioflo III, New Brunswick Scientific), in which aperforated plate was placed, was used for steeping;

[0178] temperature was only controlled during the wet stages; during theair rest stages the system was allowed to reach room temperature (ca.20° C.);

[0179] during the whole steeping period the barley was aerated (4 litersstartle air per minute);

[0180] steeping was carried out by immersion using the followingschedule: Temperature (° C.) Duration (h) First wet stage 13 6:00 Firstair rest stage 20 17:00  Second wet stage 14 5:00 Second air rest stage20 15:30  Third wet stage 16 2:30

[0181] Addition of the Microbial Cultures

[0182] 460 g of steeped barley was immersed in 0.5 liters of tap waterwhich contained no spores (A3), non-activted spores of Rhizopus oryzaeATCC 9363 (B3) or actvated spores of Rhizopus oryzae ATCC 9363 (C3according to the invention); for B3 and C3, the steeped barley wasinoculated wfth 10⁴ spores per gram of air dry barley;

[0183] the fluid was removed by draining.

[0184] Germination

[0185] as described in Example 1

[0186] Kilning

[0187] as described in Example 1

[0188] 4. Methods of analysis and results

[0189] These were as described in Example 1 (4. Methods of Analysis andResults).

[0190] See table on next page. In this table:

[0191] A1/3: Traditional malting process

[0192] B1/3: Malting process using non-activated spores

[0193] C1/3: Malting process according to the invention Example 3Example 1 pH control of the No pH control of the steeping water (pH-5.5)steeping water A3 B3 C3 A1 B1 C1 Moisture 3.8 3.6 3.7 3.9 4.1 3.8Extract 78.9 80.2 80.7 80.3 80.4 80.3 Extract 0.6 0.7 0.4 0.8 0.8 0.4Difference Color 3.2 4.2 4.4 3.3 3.3 4.1 Wort Turbidity 1 1 0.8 1.3 1.20.7 Postcoloration 5.1 7 7.2 6 6 7.3 Total Protein 10.2 10.1 10 10.110.3 10 Content Soluble Protein 4 4.4 4.8 4.1 4.4 4.8 Content KolbachIndex 39.2 43.6 48 40.6 42.7 48 Viscosity 1.52 1.53 1.52 1.57 1.52 1.52pH 6.02 5.97 5.91 6.05 6.03 5.87 Diastatic Power 348 333 355 345 349 352Whole Grains 0.2 0.2 0.1 0.3 0.3 0.1 Friability 81 81 85 83 82 83.9Homogeneity 97.6 97.8 98.9 98.5 97.9 98.6 β-glucan 190 57 40 122 108 46content Filtration 210 215 200 210 265 290 Volume Modification 84.1 85.587.4 88.2 90.5 93.4 β-glucanase 202 931 1322 214 371 683 ActivityXylanase 43 65 71 28 34 56 Activity

[0194]FIG. 3 reprsents the glucanase activity, measured according toβ-Glucazym method [Megazyme (AUSTR) Pty. Ltd.] of the malted cereals A3,B3 and C3. Malt β-glucanase activity (U/kg) was calculatd as describedin example 1. A3 was obtained by the traditional malting process with pHcontrol of the steeping water (pH=5.5). B3 resulted from the maltingprocess according to the invention with the inoculation of steepedbarley with a suspension of non-activated spores of Rhizopus oryzae ATCC9363 and with pH control of the steeping water (pH=5.6). C3 was obtainedby the malting process according to the invention with the inoculavon ofthe steeped barley with a suspension of activated spores of Rhizopusoryzae ATCC 9363 and with pH control of the steeping water (pH=5.5).

[0195] These results show the increased β-glucanase activity when the pHof the steeping water is maintained at around 5.5.

[0196]FIG. 4 gives the corresponding results for xylanase activity.These were measured according to xylszym method, Megazyme ((AUSTR) Pty.Ltd. (September 1995)). Malt xylanase activity was calculated asdescibed in Example 1.

[0197] Comparison of the β-glucanase activity obtained according toexamples 1 and 3 with the β-glucanase activity according to the state ofthe art as described in WO94/29430.

[0198] In order to compare the improved results regarding β-glucenaseactivity by the present invention, we defined the factor m as follows:$m = \frac{\beta \text{-}{glucanase}\quad {activity}\quad {of}\quad {the}\quad {treated}\quad {malt}}{\beta \text{-}{glucanase}\quad {activity}\quad {of}\quad {the}\quad {controlled}\quad {malt}}$

[0199] This factor was calculated for control malt and malted treatedwith Rhizopus oryzae ATCC 9363 as described in Examples 1 and 3 of thepresent invention.

[0200] It was also calculated for the data descrbed in WO94/29430(Example 1) where Geotrichum candidum was used.

[0201] Both as described in WO94/29430, and in the present application,β-glucanase activity was determined with the beta-glucazyme method[Megazyme (AUSTR) Pty. Ltd. (April 1993)]. Therefore, malt β-glucanaseactivity (U/kg) was calculated as 380×E(590nm)+20 and one unit ofactivity was defined as the amount of enzyme required to release onemicromole of reducing sugar equivalents per minute under the definedabove conditions.

[0202] Comparison of the Results: State of the Art Invention m m Ex. 1 mEx. 3 m Gc* 1.48 Gc* 198 B1/A1 1.73 B3/A3 4.61 C1/A1 3.19 C3/A3 6.54D1/A1 18.02

[0203] The results clearly show that the present invention provides fora more drastic increase in malt β-glucanase activity than that describedearlier (WO 94/29430).

[0204] It thus appears that it is possible to obtain malted cerealshaving a β-glucanase activity increasd by at least a factor 4 comparedto the conventional maltng process wherein the addition of microbialculture is omitted.

[0205] From FIGS. 2 and 4, it also appears that it is possible to obtainmalted cereals having a xylanase activity increased by at least a factor4 compared to conventional malting process wherein the addition ofmicrobial culture is omitted.

EXAMPLE 4

[0206] 1. Preparation of the microbial cultures

[0207] Strain

[0208] S40: Aspergillus oryzae ATCC 14156

[0209] Preparation of the spore suspension

[0210] the strain was grown on PDA (Potato Dextrose Agar, Oxoid) forapproximately 7 days at 281° C.;

[0211] the spores were harvested by flooding the culture with sterilephysiological saline (0.9%/NaCl) and by rubbing the sporulated myceliumgently with a sterile spatula;

[0212] the spore suspension was washed once with sterile physiologicalsaline (0.9% NaCl) by centrifugation (5500 rpm. Sorval type SS-34®, for15 min) and resuspended in sterile physiological saline (0.9% NaCl);

[0213] the spore density was determined microscopically using a Thomacounting chamber.

[0214] Activation of the Spore Suspension

[0215] 5×10⁷ spores were transferred into 20 ml of sterile, acidifiedTSB (Tryptic Soy Broth, Oxoid), pH=5.0 and incubated in a shaking waterbath during 3 hours (1) or 1 hour (2) at 35° C.,

[0216] 2. Cereal

[0217] Clarine barley - 1995 French harvest

[0218] 3. Process

[0219] Malts wmre made by two different malting proces:

[0220] A4. traditional malting

[0221]  (without inoculation of any spore suspension)

[0222] E4. malting process according to the invention

[0223]  (inoculation of the steeped barley during the first and thirdwet stage with a suspension of activated spores of Aspergillus oryzaeATCC 14156)

[0224] Steeping

[0225] as described in Example 1

[0226] Addition of the microbial cultures

[0227] during the steeping, 5×10³ activated spores (1) per gram air drybarley were inoculated to the water of the first wet stage and 10⁴activated spores (2) per gram air dry barley were inoculated to thewater of the third wet stage (E4);

[0228] Germination

[0229] germination of ±460 g steeped barley was carried out incylindrical containers with perfored lids at a temperature of 16°-18° C.during 4 days;

[0230] air was supplied by natural diffusion;

[0231] the containers were slowly rotated on an electronicallycontrolled roller system (Cellroll®, Tecnorama); i.e., every two hoursthe containers were rolled for 15 min at 1 rpm.

[0232] Kilning

[0233] as described in Example 1

[0234] 4. Methods of the analysis and results

[0235] These were described in Example 1 (4, Methods of Analysis andResults) Method for the determination of the acrospire length accordingto Analytica-European Brewery Convenfton (Fourth Edtion, 1987, Brsuerelund Getränke-Rundschau). 0 0-¼ ¼-½ ½-¾ ¾-1 >1 1 day germination A4 0 160 39 0 0 1 day germination E4 0 0 11 77 12 0 4 days germination A4 1 131 64 3 0 4 days germination E4 1 0 1 42 49 7

[0236] Malting Traditional Malting Process According Process (A4) to theInvention (E4) Moisture 4.3 4.0 Extract 80.9 81.1 Extract Difference 1.00.3 Color 2.8 3.2 Wort Turbidity 1.6 1.0 Postcoloration 4.8 5.4 TotalProtein Content 10.1 10.0 Soluble Protein Content 3.9 4.5 Kolbach Index38.6 44.7 Viscosity 1.57 1.48 pH 5.98 5.89 Diastatic Power 197 201 WholeGrains 1.3 0.6 Friability 81 89 Homogeneity 95.0 98.4 β-glucan Content378 132 Filtration Volume 300 310 Modification 83.9 89.8 β-glucanaseActivity 309 392 Xylanase Activity 27.82 17.62

EXAMPLE 5

[0237] 1. Preparation of the Microbial Cultures

[0238] Strains

[0239] S40: Aspergillus oryzae ATCC 14156

[0240] S46: Rhizopus oryzae ATCC 9363

[0241] Preparation of the Spore Suspensions

[0242] As described in Example 4

[0243] Activiation of the Spore Suspensions

[0244] S40:

[0245]  5×10⁷ spores were transferred into 20 ml of sterile, acidifiedTSB (Tryptic Soy Broth, Oxoid) pH=5.0 and incubated in a shaking waterduring 1 hour at 35° C.;

[0246] the activated spores were harvested by cenifugation (3500 rpm,Sorvall type SS34®, for 15 min.) and resuspended in sterilephysiological saline (0.9% NaCl).

[0247] S45

[0248] 5×10⁷ spores were transferred into 20 ml of sterile, activatedTSB (Tryptic Soy Broth, Oxoid) pH=4.0 and incubated in a shaking waterbath during 5 hours at 42° C.;

[0249] the activated spores were harvested by contrifugation (3500 rpm,Sorvall type SS34®, for 15 min.) and resuspended in sterilephysiological saline (0.9% NaCl).

[0250] 2. Cereal

[0251] Caline - 1995 French harvest

[0252] 3. Process

[0253] Setup

[0254] Malts were made by two different malting prooesm:

[0255] A5. traditional malting

[0256]  (without inoculation of any spore suspension)

[0257] F5. malting process according to the invention

[0258]  (inoculation of the steeped barley during the first wet stagewith a suspension of activated spores of Aspergillus oryzae ATCC14156and after steeping with a suspension of activated spores of Rhizopusoryzae ATCC 9363)

[0259] Steeping

[0260] As described in Example 1

[0261] Addition of the Microbial Cultures

[0262] during steeping, 10⁴ activated spores of Aspergillus oryzae ATCC14156 per gram air dry barley were inoculated to the water of the firstwet stage (F5, according to the inventon);

[0263] ±460 g of steeped barley was immersed in 0.5 liters of tap waterwhich contained no spores (A5) or activated spores of Rhizopus oryzaeATCC 9363 (F5, according to the invention); for F5 the steeped barleywas inoculated with 10⁴ activated spores per gram air dry barley;

[0264] the fluid was removed by draining.

[0265] Germination

[0266] As described in Example 4.

[0267] Kilning

[0268] As described in Example 1.

[0269] 4. Methods of Analysis and Results

[0270] These were as described in Example 1 (4. Methods of Analysis andResults).

[0271] Method for the determination of the acrospire length according toAnalytica-European Brewery Convention (Fourth Edition, 1987, Brauereiund Getränke-Rundschau). 0 0-¼ ¼-½ ½-¾ ¾-1 >1 1 day germination A5 1 153 44 1 0 1 day germination F5 0 1 21 73 5 0 4 days germination A5 0 0 029 63 8 4 days germination F5 0 0 0 13 63 24

[0272] It was noted that the use of activated spores of Aspergillusoryzae ATCC improved the malt analytical specifications.

[0273] Furthermore, it was found that during the malting process, thebarley acrospire lengths were significantly longer using the processaccording to the invention in comparison to the traditional maltingprocess. Melting Process Traditional Malting According to the Process(A5) Invention (F5) Moisture 3.9 4.2 Extract 81.4 81.8 ExtractDifference 0.9 1.1 Color 38 3.8 Wort Turbidity 1.4 1.0 Postcoloration6.9 6.4 Total Protein Content 10.1 10.2 Soluble Protein Content 4.8 5.2Kolbach Index 48.0 51.3 Viscosity 1.51 1.50 pH 5.88 5.82 Diastatic Power199 214 Whole Grains 0.8 1.1 Friability 89 95 Homogeneity 98.3 98.3β-glucan Content 120 51 Filtration Volume 270 220 Modification 96.8 98.6β-glucanase Activity 263 907 Xylanase Activity 28.86 57.76

EXAMPLE 6

[0274] 1. Preparation of the Microbial Cultures

[0275] Strains

[0276] S46: Rhizopus oryzae ATCC 9363

[0277] Preparation of the Spore Suspensions

[0278] As described in Example 4

[0279] Activation of the Spore Suspensions

[0280] 5×10⁷ spores were transferred into 20 ml of sterile, acidifiedTSB (Tryptic Soy Broth, Oxoid) pH=4.0 and incubated in a shaking waterboth during 5 hours at 42° C.;

[0281] the activated spores were harvested by centrifugation (3500 rpm,Sorvall type SS-34®, for 15 min.) and resuspended in sterilephysiological saline (0.9% NaCl).

[0282] 2. Cereal

[0283] Wheat: Mobil - 1996 Belgian harvest

[0284] 3. Process

[0285] Setup

[0286] Malts were made by two different malting processes:

[0287] A6. traditional malting

[0288]  (without inoculaton of any spore suspension)

[0289] D6. malting process according to the invention

[0290]  (inoculation of the steeped wheat during the first wet stagewith a suspension of activated spors of Rhizopus oryzae ATCC 9363)

[0291] Steeping

[0292] the steeping was carried out in a 2 kg base wfth a total water(tap water) to air ratio of 1.5:1;

[0293] use was made of 2 femnentors (Bioflo III, New BrunswickScientific), in which a perforated plate was placed;

[0294] temperature was only controlled during the wet stages; during theair rest stages, the system was allowed to reach room temperature (±20°C.);

[0295] during the whole steeping period the wheat was aerated (4 litersterile air per minute);

[0296] steeping was carried out by immersion using the following scheme:Temperature (° C.) Duration (h) First wet stage 13 6:00 First air reststage 20 16:00  Second wet stage 14 4:00 Second air rest stage 20 16:00 Third wet stage 16 2:00

[0297] Addition of the Microbial Culture

[0298] during steeping, 10⁴ activated spores of per gram air dry wheatwere inoculated to the water of the first wet stage (D6);

[0299] Germination

[0300] As described in Example 4.

[0301] Kilning

[0302] As described in Example 1.

[0303] 4. Methods of Analysis and Results

[0304] These were as dscribod in Example 1 (4. Methods of Analysis andResult). Malting Process Traditional Malting According to the Process(A6) Invention (D6) Moisture 5.5 5.4 Extract 83.6 85.5 ExtractDifference 1.0 0.6 Color 3.9 7.6 Wort Turbidity 1.4 1.4 Postcoloratlon5.8 11.5 Total Protein Content 14.0 14.8 Soluble Protein Content 4.9 9.7Kolbach Index 35.0 65.5 Viscosity 1.99 1.79 pH 6.02 5.63 Diastatic Power183 193 Whole Grains 19.4 20.2 Friability 35 42 Homogeneity 79.4 78.7Filtration Volume 220 295 β-glucanase Activity 10.9 16,640 XylanaseActivity 16.85 1,620.1

EXAMPLE 7

[0305] 1. Preparation of the Microbial Cultures

[0306] Strain

[0307] S46: Rhizopus oryze ATCC 9363

[0308] Preparation of the Spore Suspension

[0309] the strain was grown on PDA (Potato Dextrose Agar, Oxoid) forapproximately 7 days at 28° C.;

[0310] the spores were harvested by flooding the culture with sterilephysiological saline (0.9% NaCl) and by rubbing the sporulated myceliumgently with a sterile spatula;

[0311] the spore suspension was washed once with sterile physiologicalsaline (0.9% NaCl) by centifugation (3500 rpm, Jouan C312, for 15 min.)and resuspended in serile physiological saline (0.9% NaCl);

[0312] the spore density was determined microscopically using a Thomacounting chamber.

[0313] Activation of the Spore Suspension

[0314] 5×10⁷ spores were transforred into 20 ml of sterile, acidifiedTSB (Tryptic Soy Broth, Oxoid) pH=4.0 and incubated in a shaking waterbath during 5 hours at 42° C.

[0315] 2. Cereal

[0316] Sorghum (S14)

[0317] 3. Process

[0318] Setup

[0319] Malts were made by two difftrent malting processes:

[0320] A7. traditional malting

[0321]  (without inoculation of any spore suspension)

[0322] D7. malting process accordng to the invention

[0323]  (inoculation of the sorghum during the first wet stage with asuspension of activated spores of Rhizopus oryzae ATCC 9363).

[0324] Cleaning

[0325] washing of the sorghum is perforrned by using 6 liters tap waterper kilogram sorghum and by mvng the excess water.

[0326] Steeping

[0327] the steeping was carried out in a 2 kg base with a total water(top water) to air ratio of 1.5:1;

[0328] use was made of 2 fermentors (Bioflo III, New BrunswickScientific), in which a perforated plate was placed;

[0329] temperature was only controlled during the wet stages; during theair rest stages, the system was allowed to reach room temperature (±20°C.);

[0330] during the whole steeping period the barley was serated (2 litersterile air per minute);

[0331] steeping was carried out by immersion using the following scheme:Temperature (° C.) Duration (h) First wet stage 28 10:00 First air reststage 20  4:00 Second wet stage 28 10:00 Second air rest stage 20  4:00Third wet stage 28 10:00 Third air rest stage 20  4:00

[0332] Addition of the Microbial Cultures

[0333] during steeping, 10⁴ activated spores (1) per gram air drysorghum were inoculated to the water of the frst wet stage (D7).

[0334] Germination

[0335] germination of ±460 g steeped sorghum was carried out incylindrical container with perforated lids at a temperature of 28° C.during 4 days;

[0336] air was supplied by natural diffusion;

[0337] the containers were slowly rotated on an electronicallycontrolled roller system (Cellroll®, Tecnorama); i.e., every two hoursthe containers were rolled for 15 min. at 1 rpm.

[0338] Kilning

[0339] As described in Example 1.

[0340] 4. Method of Analysis and Results

[0341] These were as described in Example 1 (4. Methods of Analysis andResults). Traditional Malting Malting Process According Process (A7) tothe invention (D7) β-glucanase Activity 98 991 Xylanase Activity 524.72413.43

What is claimed is:
 1. A process for the preparation of a malted cerealcomprising the step of introducing an activated spore before or duing amalting process.
 2. The process according to claim 1 , wherein saidactivated spore increases an activity of an enzyme during said maltingprocess.
 3. The process acoording to claim 1 or claim 2 , wherein saidenzyme is selected from the group of β-glucanase, xylanase, amylase, aprotease, naturally occurring enzymes in the cereal and combinationsthereof.
 4. A process for the preparation of a malted cereal as recitedin claim 2 wherein the cereal, water and activated spores are combinedto form a combination and where the concentration of the activatedspores and the combination is held together for a time and temperaturewhich are effective for providing the malted cereal with an enzymeactivity which is greater than the enzyme activity which is obtained bya matter process without activated spores.
 5. A process as recited inclaim 4 wherein the combination is held for a time and temperature untilthe cereal has a moisture content of at least about 20 weight percent.6. A process as recited in claims 4 or 6 wherein the combination is helduntil the cereal germinates and after germination, cereal is dried to amoisture content of not more than about 15 weight percent.
 7. A processas recited in claim 6 wherein the combinaton is hold until the cerealhas a moisture content of between about 20 to about 60 weight percentand has germinated for about 2 to about 7 days at a temperature fromabout 10 to about 30° C.
 8. A process as recied in claim 6 wherein thecombination is held until the cereal has a moisture content of betweenabout 20 to about 60 weight percent and has germinated for about 2 toabout 7 days at a temperature of from about 10 to about 30° C. and isdried to a moisture content of from about 2 to about 15 weight percent.9. A process as recited in claims 1, 2 or 4 wherein the acted spores arefrom the microbes selected from the group comprising of Micrococcusspp., Streptococcus spp., Leuconostoc spp., Pediococcus spp.,Pediococcus holophilus, Pediococcus cerevisiae, Pediococcus damnosus,Pediococcus hemophilus, Pediococcus paryulus, Pedlococcus soya,Lactococcus spp., Lactobacillus spp., Lactobacillus acidophilus,Lactobacillus amylovorus, Lactobacillus bavaricus, Lactobacilusbfementens, Ladobacilus brevis var lindneri, Lactobacillus casel varcasel, Lactobacillus delbrueckii, Lactobacillus delbruecki var lacffs,Lactobaclius delbruecki var bulgericus, Lactobacillus fermenti,Lactobacillus gasserii, Lactobacillus heivetious, Lactobacillushilgardii, Lactobacillus renteril, Lactoacilius sake, Lactobacillussativorius, Lactobacillus cremoris, Lactobacillus kefir, Lactobacilluspentoolcus, Lactobacillus celloblosus, Lactobacillus bruxellensis,Lactobacillus buchneril, Lactobacillus coryneformis, Lactobacillusconfusus, Lactobacillus florentinus, Lactobacillus viridescens,Corynebacterium spp., Propionibactertum spp., Bfdobaerium spp.,Steptomyces spp., Bacillus spp., SpoioXactidiOus spp., Aoetoacter spp.,Agrobacterium spp., Alcaligenes spp., Psoeudomonas spp., Pseudomonasamylophilia, Psoudomonas aewginosa, Pseudomonas cocovenonans,Pseudomonas mexicana, Pseudomonas pseudomallei, Gluconobacter spp.,Enterobacter spp., Erwinia spp., Klebsiella spp., Proteus spp.,Ascomycota, Dothideales, Mycosphaerellaceae, Mycosphaerella spp.,Venturiaceae, Venturia spp., Eurotiales, Monascaceae, Monuscus spp.,Trichocomacase, Emericilla spp., Euroteum spp., Eupenicillium spp.,Neosartorya spp., Talaromyces spp., Hypocreales, Hypocreceae, Hypocreaspp., Saccharomycetales, Dipodascaceae, Dipodascus spp., Galactomycesspp., Endomycetaceae, Endomyces spp., Metschnikowiaceae,Guilliermondella spp., Saccharomycetaceae, Debaryomyces spp., Dekkeraspp., Pichia spp., Kluyveromyces spp., Saccharomyces spp., Torulasporaspp., Zygosaccharomyces spp., Saccharomycodaceae, Hanseniaspora spp.;Schizosaccharomycetales, Schizosaccharomycetaceae, Schizosaccharomycesspp., Sordariales, Chaetomiaceae, Chaetomium spp., Sordariacea,Neurospora spp., Zygomycota, Mucorales, Mucoraceae, Absidia spp.,Amylomyces spp., Rhizomucor spp., Actinomurco spp., Thermomucor spp.,Chiamydomucor spp., Mucor spp., Mucor circinelloides, Mucor grisecyanus,Mucor hiemalis, Mucor indicus, Mucor mucedo, Mucor piriformis, Mucorplumbeus, Mucor praini, Mucor pusillus, Mucor silvaticus, Mucorjavanicus, Mucor racernosus, Mucor rouxienus, Mucor rouxii, Mucoraromaticus, Mucor flavus, Mucor miehei, Rhizopus spp., Rhizopusarrhizus, Rhizopus oligosporus, Rhizopus oryzae, Rhizopus oryzae strainATCC 4868, Rhizopus oryzae strain ATCC 9363, Rhizopus oryzae strain NRRL1891, Rhizopus oryzae strain NRRL 1472, Rhizopus stolonifer, Rhizopusthailandensis, Rhizopus formosaensis, Rhizopus chinensis, Rhizopuscohnli, Rhizopus japonicus, Rhizopus nodosus, Rhizopus delemar, Rhizopusacetorinus, Rhizopus chlarmydosporus, Rhizopus circinans, Rhizopusjavenicus, Rhizopus peka, Rhizopus saito, Rhizopus tritici, Rhizopusniveus, Rhizopus microsporus, Mitosporic fungi, Aureobtsodium spp.,Acremonium spp., Cercospora spp., Epoocum spp., Monilia asp., Monlliacandida, Monlia sitophila, Mycoderma spp., Candida spp., Candidediddensiae, Candida edax, Candida etchellsil, Candida kefir, Candidakrisei, Candida lactose, Candida lambica, Candida melinil, Candidautilis, Candida milleri, Candida mycodema, Candida parapsilosis, Candidaobtux, Candida tropicalis, Candida valida, Candida versatilis, Candidaguilliermondii, Rhodotorula spp., Torulopsis spp., Geotrichum spp.,Geotriohum amycellum, Geotrichum armilliariae, Geotrichum asteroides,Geotrichum bipunchatum, Geotrichum dulcitum, Geotrichum eriense,Geotrichum fici, Geotrichum flavo-brunneum, Geotrichum fragrans,Geotrichum gracile, Geotrichum heritum, Geotrichum kiebaknii, Geotrichumpenicillatum, Geotrichum hirtum, Geotrichum pseudocandidum, Geotrichumrectangulatum, Geotrichum suaveoiens, Geotrichum vanrylae, Geotrichumloubieri, Geotrichum microsporum, Cladosporium spp., Trichoderma spp.,Trichoderma hamatum, Trichoderma harzianum, Trichoderma koningii,Trichoderma pseudokoningii, Trichoderma resei, Trichoderma virgatum,Trichoderma viride, Oldium spp., Altemaria spp., Altemaria altermata,Altemaria tenuis, Heiminthosporaum spp., Heiminthosporium gramineum,Helminthosporium sativum, Helminthosporium teres, Aspergillus spp.,Aspergillus ochraseus, Aspergillus nidulans, Aspergillus versicolor,Asperguillus wentil Group, Aspergilius candidus, Aspergillus flavus,Aspergillus niger, Aspergillus oryzae strain ATCC 14156, Penicillumspp., Penicillium aculeatum, Penicillum citrinum, Penicillum claviforme,Penicillum funiculosum, Penicillum itallcum, Penicillum lanoso-viride,Penicillum emersonii, Penicillum liladnum, Penicillum expansum andmixtures thereof.
 10. A malted cereal product made according to theprocess of claims 1 through
 9. 11. An aqueous combinaion of a cereal andactivated spores.
 12. A process as recited in claim 11 wherein theactivated spores are from the microbes selected from the groupcomprising Micrococcus spp., Streptococcus spp., Leuconostoc spp.,Pediococcus spp., Pediococcus halophilus, Pediococcus cerevisiae,Pediococcus damnosus, Pediococcus hemophilus, Pediococcus parvulus,Pediococcus soyae, Latococcus spp., Lactobacillus spp., Lactobacillusacidophilus, Lactobacillus amylavorus, Lactobacillus bavaricus,Lactobacillus bifermentans, Lactobacillus brevis var lindneri,Lactobacillus casel var casel, Lactobacillus delbrueckii, Lactobacillusdelbrueckii var lactis, Lactobacillus delbrueckii var bulgaricus,Lactobacillus fermenti, Lactobacillus gasserii, Lactobacillushelveticus, Lactobadlius hilgardii, Lactobacillus renteril,Lactobacillus sake, Lactobacillus sativorius, Lactobacillus cremoris,Lactobacillus kefir, Lactobacillus pentoceticus, Lactobacilluscolloblosus, Lactobacillus bruxellensfe, Lactobacillus buchnerii,Lactobacillus coryneformis, Lactobacillus confusus, Lactobacillusflorentinus, Lactobacillus viridescens, Corynebacterium spp.,Propionibacterium spp., Bifidobacterium spp., Streptomyces spp.,Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacteriumspp., Alcaligenes spp., Pseudomonas spp., Pseudomonas amylophilla,Pseudomonas aruginosa, Pseudomonas cocovenonans, Pseudomonas mexicana,Pseudomonas pseudomallel, Gluconobacter spp., Enterobacter spp., Erwiniaspp., Kiebsiella spp., Proteus spp., Ascomycota, Dothideales,Mycosphaerellaceae, Mycosphaerella spp., Venturiucoe, Venturia spp.,Eurotiales, Monaswoae, Monascus spp., Trichocomaceae, Emericilla spp.,Euroteum spp., Eupenicillium spp., Neosartorya spp., Talaromyces spp.,Hypocreales, Hypocreceae, Hypocrea spp., Saccharomycetales,Dipodascaceae, Dipodascus spp., Galactomyces spp., Endomycetaceae,Endomyces spp., Metschnikowiaceae, Guilliermondella spp.,Saccharomycetaceae, Debaryomyces spp., Dekkera spp., Pichia spp.,Kluyveromyces spp., Saccharomyces spp., Torulaspora spp.,Zygosaccharomyces spp., Saccharomycodaceae, Hanseniaspora spp.;Schizosaccharomycetales, Schizosaccharomycetaceae, Schizosaccharomycesspp., Sorderiales, Chaetomiaceae, Chaetolmium spp., Sordariacea,Neurospora spp., Zygomycota, Mucorales, Mucoraceae, Absidia spp.,Amylomyces spp., Rhizomucor spp., Actinomurcor spp., Thermomucor spp.,Chiamydomucor spp., Mucor spp., Mucor circinelloides, Mucor grisecyanus,Mucor hiemalis, Mucor indicus, Mucor mucedo, Mucor piriformis, Mucorplumbeus, Mucor praini, Mucor pusillus, Mucor silvaicus, Mucorjavanicus, Mucor racemosus, Mucor rouxianus, Mucor rouxil, Mucoraromaticus, Mucor flavus, Mucor miehei, Rhizopus spp., Rhizopusarrhizus, Rhizopus pligosporus, Rhizopus oryzae, Rhizopus oryzae strainATCC 4858, Rhizopus oryzae strain ATCC 9363, Rhizopus oryzae strain NRRL1891, Rhizopus oryzae strain NRRL 1472, Rhizopus stolonifer, Rhizopusthailandensis, Rhizopus formosaensis, Rhizopus chinensis, Rhizopuscohnii, Rhizopus japonicus, Rhizopus nodosus, Rhizopus delemar, Rhizopusacetorinus, Rhizopus chiamydosporus, Rhizopus circinans, Rhizopusjavanicus, Rhizopus peka, Rhizopus saito, Rhizopus tritici, Rhizopusniveus, Rhizopus microsporus, Mitosporic fungi, Aureobasidium spp.,Acremonium spp., Cercospora spp., Epicoccum spp., Monilia spp., Moniliacandida, Monillia sitophila, Mycoderma spp., Candida spp., Candidadiddensiae, Candida edax, Candida etchellsii, Candida kefir, Candidakrisei, Candida lactosa, Candida lambica, Candida melinli, Candidautilis, Candida millieri, Candida mycoderma, Candida parapsilosis,Candida obtux, Candida tropicalis, Candida valida, Candida versatills,Candida guilliermondii, Rhodotoruia spp., Torulopsis spp., Geotrichumspp., Geotrichum amycelium, Geotrichum armillariae, Geotrichumasteroides, Geotrichum bipunctatum, Geotrichum dulcitum, Geotrichumeriense, Geotrichum fici, Geotrichum flavo-brunneum, Geotrichumfragrans, Geotrichum gracile, Geotrichum heritum, Geotrichum kiebaknii,Geotrichum penicillatum, Geotrichum hirtum, Geotrichum pseudocandidum,Geotrichum rectangulatum, Geotrirhum suaveoiens, Geotrichum venrylae,Geotrichum loubieri, Geotrichum microsporum, Cladosporium spp.,Trichoderma spp., Trichoderma hamatum, Trichoderma harzianum,Trichoderma koningii, Trichoderma pseudokoningii, Trichoderma reesei,Trichoderma virgatum, Trichoderma viride, Oidium spp., Altemaria spp.,Altemaria altemata, Altemaria tanuis, Helminthosporium spp.,Helminthosporium gramineum, Helminthosporium sativum, Helminthosporiumteres, Aspergillus spp., Aspergillus ochraseus, Aspergillus nidulans,Aspergillus versicolor, Aspergillus wentii Group, Aspergillus candidus,Aspergillus flavus, Aspergillus niger, Aspergillus oryzae strain ATCC14156, Penicillum spp., Penicillum aculeatum, Penicillum citrinum,Penicillum claviforma, Penicillum funiculosum, Penicillum italicum,Penicillum lanoso-viride, Penicillum emersonii, Penicillum lilacinum,Penicillum expansum and mixtures thereof.
 13. A process for thepreparation of a malted cereal said process comprising the steps of: (a)introducing an activated spore into a moistened cereal to provide aninoculated moistened cereal to form a moistened cereal/acivated sporecombination; (b) germinating said inoculated moistened cereal; and (c)drying said germinated cereal.
 14. The process according to claim 13 ,wherein said inoculated moistened cereal is held at a temperature offrom about 5° to about 30° C. until the cereal has a moisture content offrom about 20 to about 60 weight percent moisture.
 15. The processaccording to claim 13 or claim 14 , wherein said germinating step (b) iscarried out for about 3 to about 6 days at a temperature of from about10° to about 30° C.
 16. The process according to any one of claims 13 to15 , wherein said germinated cereal is dried to a moisture content offrom about 2 io about 15 weight percent.
 17. A process as recited inclaim 13 wherein the combination is held at a temperature of from about10° C. to about 20° C. until the cereal has a moisture content of fromabout 38 to about 47 weight percent and the cereal has germinated forabout 3 to about 6 days at a temperare of from about 14° C. to about 18°C. and the germinated cereal is dried at a temperature of from about 40°C. to about 150° C.
 18. A process for the preparation of a malted cerealsaid process comprising the step of moistening a cereal and activatedspores wherein the concentration of the activated spores, moisteningtime and moistening temperature are effective for providing the maltedcereal with an increase in activity of an enzyme compared to theactivity of an enzyme obtained by moistening the cereal withoutactivated spores.
 19. The process according to claim 18 , wherein, saidenzyme is selected from the group of β-glucanase, xylanase, amylase,protease, naturally occurring enzymes in the cereal and combinationsthereof.
 20. A process as recited in claim 18 wherein the cerealmoistening time and temperature are effective to provide the cereal witha moisture content of at least about 20 weight percent.
 21. A process asrecited in claim 20 wherein after cereal attains a moisture content ofat least about 20 weight percent, it is dried to a moisture content ofnot more than about 15 weight percent.
 22. A process as recited inclaims 18, 19, 20 or 21 wherein the moistening time and temperature areeffective to provide the cereal with a moisture content of between about20 to about 60 weight percent and wherein the cereal has germinated forabout 2 to about 7 days at a temperature of from about 10 to about 30°C.
 23. A process as recited in claim 22 wherein the germinated is driedto a moisture content of from about 2 to about 15 weight percent.
 24. Aprocess as recited in claim 23 wherein the activated spores are from themicrobes selected from the group comprising Micrococcus spp.,Streptococcus spp., Leuconostoc spp., Pediococcus spp., Pediococcushalophilus, Pediococcus cerevisiae, Pediococcus damnosus, Pediococcushemophilus, Pediococcus parvulus, Pediococcus soyae, Lactococcus spp.,Lactobacillus spp., Lactobacillus acidophilus, Lactobacillus amylovorus,Lactobacillus bavaricus, Lactobacillus bifermentans, Lactobacillusbravis var lindneri, Lactobacillus casel var cesi, Lactobacillusdelbrueckii, Lactobacillus delbrueckii var lactis, Lactobacillusdelbrunckii var bulgaricus, Lactobacillus fermenti, Lactobacillusgasserii, Lactobacillus helveticus, Lactobacillus hilgardii,Lactobacillus renteril, Lactobacilus sake, Lactobacilus sativorius,Lactobacillus cremoris, Lactobacillus kefir, Lactobacillus pentoceticus,Lactobacillus cellobiosus, Lactobacillus bruxellensis, Lactobacillusbuchnerii, Lactobacillus coryneformis, Lactobacillus confusus,Lactobacillus florentinus, Lactobacillus viridescens, Corynebaceriumspp., Propionibacterium spp., Bifidobacterium spp., Streptomyces spp.,Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacteriumspp., Alcaligenes spp., Pseudomonas spp., Pseudomonas amylophilia,Pseudomonas aeruginosa, Pseudomonas cocovenenas, Pseudomonas mexicana,Pseudomonas pseudomallei, Gluaonobacter spp., Enterobacter spp., Erwiniaspp., Kebsiella spp., Proteus spp., Ascomycota, Dothideales,Mycosphaerellaceae, Mycosphaerella spp., Venturiaceae, Venturia spp.,Eurotiales, Monascaceae, Monascus spp., Trichocomaceae, Emericilla spp.,Euroteum spp., Eupenicillium spp., Neosartorya spp., Talaromyces spp.,Hypocreales, Hypocreaceae, Hypocrea spp., Saccharomycetales,Dipodascaceae, Dipodascus spp., Galactomyces spp., Endomycetaceae,Endomyces spp., Metschnikowfaceae, Guilliermondella spp.,Saccharomycetaceae, Debaryomyces spp., Dekkera spp., Pichia spp.,Kluyveromyces spp., Saccharomyces spp., Torulaspora spp.,Zygosaccharomyces spp., Saccharomycodacea, Hanseniaspora spp.;Schizosaccharomycetales, Schizosaccharomycetaceae, Schizosaccharomycesspp., Sordariales, Chaetomiaceae, Chaetomium spp., Sorderiacea,Neurospora spp., Zygomycota, Mucorales, Mucoraceae, Absidia spp.,Amylomyces spp., Rhizomucor spp., Actinomucor spp., Thermomucor spp.,Chiamydomucor spp., Mucor spp., Mucor circinelloides, Mucor grisecyanus,Mucor hiemalls, Mucor indicus, Mucor mucedo, Mucor piriformis, Mucorplumbeus, Mucor praini, Mucor pusillus, Mucor slivaticus, Mucorjavanicus, Mucor racemosus, Mucor rouxianus, Mucor rouxii, Mucoraromaticus, Mucor flavus, Mucor miehei, Rhizopus spp., Rhizopusarrhizus, Rhizopus olgosporus, Rhizopus oryzae, Rhizopus oryzae strainATCC 4858, Rhizopus oryzae strain ATCC 9303, Rhizopus oryzae strain NRRL1891, Rhizopus oryzae strain NRRL 1472, Rhizopus stolonifer, Rhizopusthailandensis, Rhizopus formosaensis, Rhizopus chinensis, Rhizopuscohnli, Rhizopus japonicus, Rhizopus nodosus, Rhizopus delemar, Rhizopusacetorinus, Rhizopus chlamydosporus, Rhizopus circinans, Rhizopusjavanicus, Rhizopus peka, Rhizopus saito, Rhizopus tritici, Rhizopusniveus, Rhizopus microsporus, Mitosporic fungi, Aureobasidium spp.,Acremonium spp., Cerspora spp., Epicoccum spp., Monllia spp., Monillacandida, Monilia sitophila, Mycoderma spp., Cendida spp., Candidadiddensiae, Candida edex, Candida etcheilsil, Candida kefir, Candidakrisei, Candida lactosa, Candida lambica, Candida melinli, Candidautilis, Candida milleri, Candida mycoderma, Candida parapsilosis,Candida obtux, Candida tropicalis, Candida valida, Candida versatilis,Candida guilliermondil, Rhodatorula spp., Torulopsis spp., Geotrichumspp., Geotrichum amycellium, Goetrichum armillariae, Geotrichumasteroides, Geotrichum bipunctatum, Goetrichum dulcitum, Geotrichumeriense, Geotrichum fici, Geotrichum flavo-brunneum, Geotrichumfragrans, Geotrichum gracile, Geotrichum heritum, Geotrichum kiebaknii,Geotrichum penicillatum, Geotrichum hirtum, Geotrichum pseudocandidum,Geotrichum rectangulatum, Geotrichum suaveolens, Geotrichum vanryiae,Geotrichum loubieri, Geotrichum microsporm, Cladosporium spp.,Trichoderma spp., Trichodenna hamatum, Trichoderma harzienum,Trichoderma koningii, Trichoderma pseudokoningii, Trichoderma reesei,Trichoderma virgatum, Trichoderma viride, Oldium spp., Altemaria spp.,Altemaria altermata, Altemaria tenuis, Helminthosporium spp.,Helminthosporium gramineum, Helminthosporium sativum, Helminthosporiumteres, Aspergillue spp., Aspergillus ochraseus, Aspergillus nidulans,Aspergillus versicolor, Aspergilius wentil Group, Aspergillus candidus,Aspergillus flavus, Aspergillus niger, Aspergillus oryzae strain ATCC14156, Penicillum spp., Penicilum aculeatum, Penicillum citrinum,Penicillum claviforme, Penicillum funiculosum, Penicillum italicum,Penicillum lanoso-viride, Peniciflum emersonii, Penicilium lilacinum,Penicillum expensum and mixtures thereof.
 25. A malted cereal productmade according to the proces of claims 18, 19, 20, 21, 22, 23 or
 24. 26.Use of activated spores in the preparation of a malted cereal.
 27. Aprocess for the preparation of malted cereals, wherein the steeping stepincludes one or morm wetting stages at a temperature between 5° and 30°C., preferbly between 10° and 20° C., until the material has a moisturecontent between 20% and 60% by weight, preferably between 38% and 47%,wherein after a germination period between 2 and 7 days, preferablybetween 3 to 6 days at a temperature between 10° and 30° C., preferablybetween 14° and 18° C., the steeped and germinated cereals arepreferably kilned by increasing the temperature to values 40° and 150°C. until the material has a moisture content between 2% and 15% byweight, and wherein one or more microbial cultures selected from thegroup consisting of one o more bacteria and/or one of more fungi areadded in one or more times either before or during or after the maltingprocess of said cereals.
 28. Pass according to claim 27 , for thepreparation of malted barley, wherein the bacteria are selected from thegroup comprising Micrococcus spp., Streptococcus spp., Leuconostoc spp.,Pediococcus spp. preferentially Pediococcus halophilus, Pediococcuscerevisiae, Pediococcus damnosus, Pediococcus hemophilus, Pediococcusparvulus, Pediococcus soyae, Lactococcus spp., Lactobacius spp.preferentially Lactobacillus acidophilus, Lactobacillus amylovorus,Lactobacillus bavaricus, Lactobacillus bifermentens, Lactobacillusbrevis var lindneri, Lactobacillus casel var easel, Lactobacillusdelbrueckil, Lactobacillus delbrueckii var lactis, Lactobacillusdelbrueckil var bulgaricus, Lactobacillus fermenti, Lactobacillusgasserii, Lactobacillus helveticus, Lactobacillus hilgardil,Lactobacillus renteril, Lactobacillus sake, Lactobacillus satvorius,Lactobacilus cremoris, Lactobachillus kefir, Lactobacillus pentoceticus,Lactobacillus celloblosus, Laclocilus bruxellensis, Lactobacillusbuchnerii, Lactobacillus coryneformis, Lactobacillus confusus,Lactobacillus florentinus, Lactobacillus viridescens, Corynebacteriumspp., Propionibacterium spp., Bifidobacterium spp., Streptomyces spp.,Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacierimspp., Alcallgenes spp., Pseudomonas spp. preferentially Pseudomonasamylophillis, Pseudomonas aeruginosa, Pseudomonas cocovenenana,Pseudomonas mexicana, Pseudomonas pseudomallei, Gluconobacter spp.,Enterobacter spp., Erwinia spp., Kiebsiella spp., and Proteus spp. 29.The prooess according to claim 27 , for the preparation of malted barleywherein the fungi are selected from the group (geners as described byAinsworth and Bisby's dictionary of the fungi, 8th edition, 1995, editeby D. L. Hawksworth, P. M. Kirk, B. C. Sutton, and D. N. Pegler (632 pp)Cab Inbemational) comprising Ascomycota preferentially Dothidealespreferentially Mycosphaerellaceae preferentially Mycosphaerella spp.,Venturfaceae preferentially Venturia spp.; Eurotiales preferentiallyMonascaceae preferentially Monascus spp., Trichocomaceae preferentiallyEmericilla spp., Euroteum spp., Eupenicillium spp., Neosartorya spp.,Talaromyces spp.; Hypocreales preferentially Hypocreceae preferentiallyHypocrea spp.; Saccharomycetales preferentially Dipodascaceaepreferentially Dipodacus spp., Galactomyces spp., Endomycetaceaepreferentially Endomyces spp., Metschnikowiaceae preferentiallyGuilliermondella spp., Saccharomycetaceae preferentially Debaryomycesspp., Dekkera spp., Pichia spp., Kluyveromyces spp., Saccharomyces spp.,Torulaspora spp., Zygosaccharomyces spp., Saccharomycodaceaepreferentially Hanseniaspora spp.; Schlzosaccharomycetalespreferentially Schizosaccharomycetaceae preferentiallySchizosaccharomyces spp.; Sordariales preferentially Chaetomisceaepreferentially Chaetomium spp., Sordaisceae preferentially Neursporespp.; Zygomycota preferentially Mucorales preferentiaily Mucoraceaepreferentially Absidia spp., Amylomyces spp., Rhizomucor spp.,Actinomucor spp., Thermomucor spp., Chiamydomucor spp., Mucor spp.preferentially Mucor circinelloides, Mucor grisecyanus, Mucor hiermalis,Mucor indicus, Mucor mucedo, Mucor piriformis, Mucor plumbeus, Mucorpraini, Mucor pusillus, Mucor silvaticus, Mucor javanicus, Mucorracemosus, Mucor rouxianus, Mucor rouxii, Mucor aromaticus, Mucorflavus, Mucor mishel, Rhizopus spp. preferentally Rhizopus arrhizus,Rhizopus oligosporus, Rhizopus oryzae preferentially strains ATCC 4858,ATCC 9363, NRRL 1891, NRRL 1472, Rhizopus stolonifer, Rhizopusthailandensis, Rhizopus formmosaensis, Rhizopus chinensis, Rhizopuscohnii, Rhizopus japonicus, Rhizopus nodosus, Rhizopus delemar, Rhizopusacetorinus, Rhizopus chlamydosporus, Rhizopus circinans, Rhizopusjavanicus, Rhizopus peka, Rhizopus saito, Rhizopus tritici, Rhizopusniveus, Rhizopus miorospous; Mitosporic fungi preferentiallyAureobasidium spp., Acremonium spp., Cercospora spp., Epicoccoum spp.,Monilia spp. preferentially Monilia candida, Monilia sitophila,Mycoderma spp., Candida spp. preferentially Candida diddensiae, Candidaedax, Candida etchellsli, Candida kefir, Candida krisel, Candidalactose, Candida lambica, Candida melinii, Candida utilis, Candidamilleri, Candida mycoderma, Candida parapsilosis, Candida obtux, Candidatropicalis, Candida valida, Candida versatilis, Candida guilliermondii,Rhodotorula spp., Torulopsis spp. Geotrichum spp. preferentiallyGeotrichum amycefium, Geotrichum armillariae, Geotrichum asteroides,Geotrichum bipunctatum, Geotrichum dulcitum, Geotrichum eriense,Geotrichum fici, Geotrichum flavo-brunneum, Geotrichum fragrans,Geotrichum gracile, Geotrichum heritum, Geotrichum kiebaknil, Geotrichumpenicillatum, Geotrichum hirtum, Geotrichum pseudocandidum, Geotrichumrectanulatum, Geotrichum suaveolens, Geotrichum vanrylae, Geotrichumloubieri, Geotrichum micrsporum, Cladosporium spp., Trichoderma spp.preferentially Trichoderma hamatum, Trichoderma harzianum, Trichodermakoningil, Trichoderma pseudokoningii, Trichoderma reesei, Trichodermavirgatum, Trichoderma viride, Oldium spp., Altemaria spp. preferentiallyAltemaria altemata, Altemaria tenuls, Helminthosporium spp.preferentialy Helminthosporium gramineum, Holminthosporium sativum,Helminthosporium teres, Aspergillus spp. as described by R. A. Samson((1994) in Biotehnologicl handbooks, Volume 7:Aspergillus, edited bySmith, J. E. (273 pp), Plenum Press) preferentially Aspergillusochraseus Group (Thom & Churh), Aspergillus nidulans Group (Thom &Church), Aspergillus versicolor Group (Thom & Church) Aspergillus wentilGroup (Thom & Raper), Aspergillus candidus Group (Thom & Raper,Aspergillus flavus Group (Raper & Fennell), Aspergilius niger Group(Thom & Church), Penicillum spp. preferentially Penicillum aculeatum,Penicillum citrinum, Penicillum claviforme, Penicillum funiculosum,Penicillum italicum, Penicillum lanoso-viride, Penicillum amersonil,Penicillum lilacinum, and Penicillum expansum.
 30. The process accordingto claim 27 for the preparation of malted cereals other than maltedbarley wherein the bacteria are selected from the group comprisingMicrococcus spp., Streptococcus spp., Leuconostoc spp., Pediococcusspp., Lactococcus spp., Lactobacillus spp., Corynebacterium spp.,Propionibacterium spp., Bifidobacterium spp., Streptomyces spp.,Bacillus spp., Sporolactobacillus spp., Acetobacter spp., Agrobacteriumspp., Alcallgenes spp., Pseudomonas spp., Gluconobacter spp.,Enterobacter spp., Erwinia spp., Kiebsiella spp., and Proteus spp. 31.Process according to claim 27 for the preparation of malted cerealsother than malted barley wherein the fungi are selected from the groupcomprising Ascomycota preferentially Dothideales preferentiallyMycosphaerellaceae preferentially Mycosphaerella spp., Venturiaceaepreferably Venturia spp.; Eurotiales preferentially Monascaceaepreferentially Monascus spp., Trichocomaceae preferentially Emericillaspp., Euroteum spp., Eupenicillium spp., Neosartorya spp., Talaromycesspp.; Hypocreales preferentially Hypocreceae preferentially Hypocreaspp.; Saccharomycetales preferentially Dipodascaceae preferentiallyDipodascus spp., Galactomyces spp., Endomycetaceae preferentiallyEndomyces spp., Metschnikowiaceae preferentially Guilliermondella spp.,Saccharomycetaceae preferentially Debaryomyces spp., Dekkera spp.,Pichia spp., Kluyveromyces spp., Saccharomyces spp., Torulespora spp.,Zygosaccharomyces spp., Saccharomycodaceae preferentially Hanseniasporaspp.; Schizosaccharomycetales preferentially Schizosaccharomycetacaepreferentially Schizosaccharomyces spp.; Sordariales preferentiallyChaetomiaceae preferentially Chaetomium spp., Sordariacesepreferentially Neurospora spp.; Zygomycota preferentially Mucoralespreferentially Mucoracese preferentially Absidia spp., Amylomyces spp.,Rhizomucor spp., Actinomucor spp., Thermomucor spp., Chiamydomucor spp.,Mucor spp., Rhizopus spp.; Mitosporic fungi preferentially Aureobasidumspp., Acremonium spp., Cerocospora spp., Epicoccum spp., Monilia spp.,Mycoderma spp., Candida spp., Rhodotorula spp., Torulopsis spp.,Geotrichum spp., Cladosporium spp., Trichoderma spp., Oldium spp.,Altemaria spp., Helminthosporium spp., Aspergillus spp., and Penicilliumspp.
 32. Process according to any of claims 27 to 31 , wherein the totaltime of submersion in water during steeping for physiological reasonsdoes not exceed 30 hours, preferentially takes 10 to 25 hours, orwherein the kilning includes more than two temperature steps and whereinthe microbial culture comprises Rhizopus spp. and/or Pseudomonas spp.33. Process according to the claim 32 , wherein the Rhizopus spp. ispreferably a Rhizopus orzyae such as a Rhizopus oryzae strain ATCC 9363.34. Process according to the claim 31 or claim 32 , wherein thePseudomonas sp. is preferably a Pseudomonas herbicola.
 35. Processaccording to any of claims 27 to 35 , wherein the mirobial spores usedare activated by one or a combination of the foliowing treatments: (a)cycles of wotng and/or drying, (b) addition of nutritional supplies oraddition of spore elements. (c) exposure to temperature changes,preferably within a range of 0° to 80° C., (d) exposure to change in pH,preferably within a pH range of 2.0 to 8.0, more preferably between 3.0and 6.0, to obtain spores significantly more swollen than their dormantsize, more particularly, the size of the spores is increased by a factorpreferably between 1.2 and 10 over their domant size and/or spores withone or more germ tubes per spore.
 36. Process according to any one ofclalms 27 to 35, wherein the pH during the steeping step is adjusted toa value between 4.0 and 6.0.
 37. Process according to any one of claims27 to 36 , wherein nutrients and/or additives are added prior to and/orduring the malting process.
 38. Malted barley characbried by aβ-glucanase activity increased by at least a factor 4 and a xylanaseactivity increased by at least a factor of 4, compared to theconventional malting process of any available barley.
 39. Malted barley,wherein the β-glucanase activity is higher than 700 units/kg, and thexylanase activity is higher than 250 units/kg.
 40. Malted barleyaccording to claim 38 or 39 obtained by the process of any one of theclaims 27 to 37 .
 41. Malted barley according to any one of claims 36 to40 , characterized in that they present an improved modificaton and/oran increased hydrolytic enzyme activity, a decreased level of toxinsand/or increased microbial safety or increased acceptability.
 42. Use ofthe malted cereals according to any one of the claims 38 to 41 , orobtained by the process of any one of the claims 27 to 37 for thepreparation of beverages.